{"doi":"10.1172/jci155621","title":"An optimized bicistronic chimeric antigen receptor against GPC2 or CD276 overcomes heterogeneous expression in neuroblastoma","abstract":"Chimeric antigen receptor (CAR) T cell therapies targeting single antigens have performed poorly in clinical trials for solid tumors due to heterogenous expression of tumor-associated antigens (TAAs), limited T cell persistence, and T cell exhaustion. Here, we aimed to identify optimal CARs against glypican 2 (GPC2) or CD276 (B7-H3), which were highly but heterogeneously expressed in neuroblastoma (NB), a lethal extracranial solid tumor of childhood. First, we examined CAR T cell expansion in the presence of targets by digital droplet PCR. Next, using pooled competitive optimization of CAR by cellular indexing of transcriptomes and epitopes by sequencing (CITE-Seq), termed P-COCC, we simultaneously analyzed protein and transcriptome expression of CAR T cells to identify high-activity CARs. Finally, we performed cytotoxicity assays to identify the most effective CAR against each target and combined the CARs into a bicistronic \"OR\" CAR (BiCisCAR). BiCisCAR T cells effectively eliminated tumor cells expressing GPC2 or CD276. Furthermore, the BiCisCAR T cells demonstrated prolonged persistence and resistance to exhaustion when compared with CARs targeting a single antigen. This study illustrated that targeting multiple TAAs with BiCisCAR may overcome heterogenous expression of target antigens in solid tumors and identified a potent, clinically relevant CAR against NB. Moreover, our multimodal approach integrating competitive expansion, P-COCC, and cytotoxicity assays is an effective strategy to identify potent CARs among a pool of candidates.","journal":"Journal of Clinical Investigation","year":2022,"id":236141,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":72,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.956,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":855947,"name":"Adam Cheuk","orcid":"0000-0001-6327-1631","position":1,"is_corresponding":false},{"id":250438,"name":"Jun S. Wei","orcid":"0000-0002-4812-0250","position":2,"is_corresponding":false},{"id":425135,"name":"Abdalla Abdelmaksoud","orcid":null,"position":3,"is_corresponding":false},{"id":506189,"name":"Hsien-Chao Chou","orcid":"0000-0002-4870-9663","position":4,"is_corresponding":false},{"id":347040,"name":"David Milewski","orcid":"0000-0002-9778-8548","position":5,"is_corresponding":false},{"id":226280,"name":"Michael C. Kelly","orcid":"0000-0003-0654-2778","position":6,"is_corresponding":false},{"id":855948,"name":"Young Song","orcid":"0000-0002-0035-8649","position":7,"is_corresponding":false},{"id":310940,"name":"Christopher M. Dower","orcid":"0000-0003-2068-6524","position":8,"is_corresponding":false},{"id":255115,"name":"Nan Li","orcid":"0000-0002-1498-7139","position":9,"is_corresponding":false},{"id":309021,"name":"Haiying Qin","orcid":"0000-0001-5966-8812","position":10,"is_corresponding":false},{"id":670843,"name":"Yong Kim","orcid":"0000-0002-5417-9232","position":11,"is_corresponding":false},{"id":856804,"name":"Jerry T. Wu","orcid":null,"position":12,"is_corresponding":false},{"id":648067,"name":"Xinyu Wen","orcid":"0000-0001-8233-5418","position":13,"is_corresponding":false},{"id":843713,"name":"Mehdi Benzaoui","orcid":"0000-0003-4453-367X","position":14,"is_corresponding":false},{"id":41980,"name":"Katherine  E. Masih","orcid":"0000-0002-0809-668X","position":15,"is_corresponding":false},{"id":255121,"name":"Xiaolin Wu","orcid":"0000-0002-6432-1300","position":16,"is_corresponding":false},{"id":855949,"name":"Zhongmei Zhang","orcid":"0000-0001-7450-8040","position":17,"is_corresponding":false},{"id":856805,"name":"S. Badr","orcid":null,"position":18,"is_corresponding":false},{"id":113313,"name":"Naomi Taylor","orcid":"0000-0002-2459-4558","position":19,"is_corresponding":false},{"id":275780,"name":"Brad St. Croix","orcid":"0000-0002-6246-0374","position":20,"is_corresponding":false},{"id":255131,"name":"Mitchell Ho","orcid":"0000-0002-9152-5405","position":21,"is_corresponding":false},{"id":250453,"name":"Javed Khan","orcid":"0000-0002-5858-0488","position":22,"is_corresponding":false},{"id":555396,"name":"Meijie Tian","orcid":"0000-0003-0547-9447","position":0,"is_corresponding":true}],"reference_count":53,"raw_metadata":null,"created_at":"2026-07-19T00:21:58.728756Z","pmid":"35852863","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}